Anti-TOR1AIP1 antibody [EPR29003-34]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling TOR1AIP1 with ab317701 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling TOR1AIP1 with ab317701 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nucleus envelope staining on human tonsil.
The section was incubated with ab317701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Immunohistochemical analysis of paraffin-embedded Human melanoma tissue labeling TOR1AIP1 with ab317701 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nucleus envelope staining on human melanoma (PMID : 36624187).
The section was incubated with ab317701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling TOR1AIP1 with ab317701 at 1/2000 (0.249 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nucleus envelope staining on human testis (PMID : 28387711).
The section was incubated with ab317701 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling TOR1AIP1 with ab317701 at 1/500 (0.994 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing nuclear envelope staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• WB

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Western blot - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Lanes 1-3 were freshly made and used for Western Blotting immediately to minimize protein degradation.

Lanes 1-3 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 4-5 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/5000.

The identity of the band at ~30KDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TOR1AIP1 antibody [EPR29003-34] (ab317701) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 3:

HeLa transfected with siRNA specifically targeti TOR1AIP1 whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Lane 5:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 6:

SK-MEL-28 (human malignant melanoma cell) whole cell lysate at 20 µg

Secondary

Lanes 1 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 4 - 5:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/5000 dilution

Observed band size: 66 kDa,36 kDa

false

Exposure time: 48s

• Dot

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Dot Blot - Anti-TOR1AIP1 antibody [EPR29003-34] (AB317701)

Dot blot analysis of TOR1AIP1 using ab317701 at 1 : 1000 (0.497 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This antibody does not cross-react with human TOR1AIP2.

All lanes:

Dot Blot - Anti-TOR1AIP1 antibody [EPR29003-34] (ab317701) at 1/1000 dilution

Lane 1:

His-tagged human TOR1AIP1 fragment

Lane 2:

His-tagged human TOR1AIP2 fragment

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s